CAJ | 학술논문

背景前期研究发现,激肽原1是增生性玻璃体视网膜病变(PVR)患者玻璃体中的特异蛋白质,是在质谱鉴定中得到的肽段匹配和MASCOT得分最高的蛋白质,且与PVR的严重程度呈正相关.目的探讨激肽原-激肽系统(KKS)是否参与PVR的发生过程.方法大鼠视网膜色素上皮(RPE)细胞系RPE-J细胞复苏培养后用PBS配制成2.5×108/ml的细胞悬液,大鼠下腔静脉采血4ml经枸橼酸二钠处理和离心后用PBS制备成血小板密度为2.5×108/ml的血浆.用随机数字表法将60只Wistar大鼠随机分为实验组和生理盐水组,每组各30只.实验组大鼠左眼玻璃体腔内注射RPE细胞悬液4μl+富含血小板血浆6μl建立PVR模型,生理盐水组左眼玻璃体腔以同样的方法注射生理盐水10μl,各组大鼠的右眼作为自身对照组.术后1、3、7、14、21、28d行裂隙灯及间接检眼镜检查,按Francine的标准进行PVR分级.玻璃体腔注射后28d收集实验动物的玻璃体、血清和视网膜标本,应用Western blot法检测缓激肽的表达,视网膜标本行组织病理学检查.结果术后28d,实验组有25只眼出现不同程度的PVR表现,建模成功率为89.3%(25/28).术后7、14、28d裂隙灯下证实实验组大鼠形成1、2、3级PVR.视网膜组织病理学检查表明视网膜组织中炎性细胞浸润及RPE细胞移行并转化为成纤维细胞,出现视网膜脱离.Western blot分析显示在所有PVR大鼠血清、玻璃体和视网膜中均可检测到缓激肽的表达,但实验组大鼠的表达强度明显高于生理盐水组大鼠.结论玻璃体腔注射RPE细胞联合富含血小板血浆的方法可以成功建立PVR大鼠模型,KKS可能参与PVR的发生.
배경 전기연구발현,격태원1시증생성파리체시망막병변(PVR)환자파리체중적특이단백질,시재질보감정중득도적태단필배화MASCOT득분최고적단백질,차여PVR적엄중정도정정상관.목적 탐토격태원-격태계통(KKS)시부삼여PVR적발생과정.방법 대서시망막색소상피(RPE)세포계RPE-J세포복소배양후용PBS배제성2.5×108/ml적세포현액,대서하강정맥채혈4ml경구연산이납처리화리심후용PBS제비성혈소판밀도위2.5×108/ml적혈장.용수궤수자표법장60지Wistar대서수궤분위실험조화생리염수조,매조각30지.실험조대서좌안파리체강내주사RPE세포현액4μl+부함혈소판혈장6μl건립PVR모형,생리염수조좌안파리체강이동양적방법주사생리염수10μl,각조대서적우안작위자신대조조.술후1、3、7、14、21、28d행렬극등급간접검안경검사,안Francine적표준진행PVR분급.파리체강주사후28d수집실험동물적파리체、혈청화시망막표본,응용Western blot법검측완격태적표체,시망막표본행조직병이학검사.결과 술후28d,실험조유25지안출현불동정도적PVR표현,건모성공솔위89.3%(25/28).술후7、14、28d렬극등하증실실험조대서형성1、2、3급PVR.시망막조직병이학검사표명시망막조직중염성세포침윤급RPE세포이행병전화위성섬유세포,출현시망막탈리.Western blot분석현시재소유PVR대서혈청、파리체화시망막중균가검측도완격태적표체,단실험조대서적표체강도명현고우생리염수조대서.결론파리체강주사RPE세포연합부함혈소판혈장적방법가이성공건립PVR대서모형,KKS가능삼여PVR적발생.
Background Our previous study demonstrated that kallikrein-kinin is a special protein in vitreous of the eye with proliferative vitreoretinopathy(PVR),and the expression intensity of kallikrein-kinin showed the positive correlation with the grade of PVR.Objective This study was to further explore whether kallikrein-kinin participate in the formation of PVR.Methods Rat retinal pigment epithelial cell line(RPE-J cells) was cultured in DMEM containing 4% fetal bovine serum and then prepared into suspension by PBS with the cells density of 2.5×108 cells/ml.Platelet-rich plasma was prepared by PBS with the platelet 2.5×108 /ml.RPE cell suspension(4μl) and platelet-rich plasma(6μl) was intravitreally injected in the left eyes of 30 clean Wistar rats to establish the PVR models,and 10μl sterile pyrogen-free normal saline solution was used in the same way in other matched rats as controls.The PVR was graded on Francine's criteria in 1 day,3,7,14,21,28 days after injection under the slit lamp.The serum,vitreous and retina were obtained in 28 days after injection to assess the expression of bradykinin using Western blot.The histopathology examination of rat retina was performed in the 28th day after injection.This experimental procedure followed the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results Typical PVR was seen in 25 models with the successful rate 89.3% at 28 days after injection.PVR 1,2,3 grades were respectively exhibited in 7,14,28 days under the slit lamp.Infiltration of inflammatory cells and migration of RPE cells were found in the 7th day.In the 14th day after injection,RPE cells transformed into fibroblasts and retinal detachment occurred after that.Western blot analysis revealed that bradykinin was detected in vitreous,serum and retinal samples of rats in experimental and control rats,but the expression intensity was higher in the rats of model groups.Conclusion Intravitreal co-injection of RPE cells and platelet-rich plasma can effectively induce a model of PVR in Wistar rat.The kallikrein-kinin system probably takes part in the onset of PVR.